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Mol Cell Proteomics. 2010 May;9(5):824-37. doi: 10.1074/mcp.M900569-MCP200. Epub 2010 Feb 4.

High resolution electron transfer dissociation studies of unfractionated intact histones from murine embryonic stem cells using on-line capillary LC separation: determination of abundant histone isoforms and post-translational modifications.

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  • 1Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94158-2517, USA.

Abstract

Epigenetic regulation of chromatin is dependent on both the histone protein isoforms and state of their post-translational modifications. The assignment of all post-translational modification sites for each individual intact protein isoform remains an experimental challenge. We present an on-line reversed phase LC tandem mass spectrometry approach for the separation of intact, unfractionated histones and a high resolution mass analyzer, the Orbitrap, with electron transfer dissociation capabilities to detect and record accurate mass values for the molecular and fragment ions observed. From a single LC-electron transfer dissociation run, this strategy permits the identification of the most abundant intact proteins, determination of the isoforms present, and the localization of post-translational modifications.

PMID:
20133344
[PubMed - indexed for MEDLINE]
PMCID:
PMC2871417
Free PMC Article

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